A single-cell atlas of chromatin accessibility in the human genome.

Current catalogs of regulatory sequences in the human genome are still incomplete and lack cell type resolution. To profile the activity of gene regulatory elements in diverse cell types and tissues in the human body, we applied single-cell chromatin accessibility assays to 30 adult human tissue types from multiple donors. We integrated these datasets with previous single-cell chromatin accessibility data from 15 fetal tissue types to reveal the status of open chromatin for ∼1.2 million candidate cis-regulatory elements (cCREs) in 222 distinct cell types comprised of >1.3 million nuclei. We used these chromatin accessibility maps to delineate cell-type-specificity of fetal and adult human cCREs and to systematically interpret the noncoding variants associated with complex human traits and diseases. This rich resource provides a foundation for the analysis of gene regulatory programs in human cell types across tissues, life stages, and organ systems.

Transcription Factors Activate Genes through the Phase-Separation Capacity of Their Activation Domains.

Gene expression is controlled by transcription factors (TFs) that consist of DNA-binding domains (DBDs) and activation domains (ADs). The DBDs have been well characterized, but little is known about the mechanisms by which ADs effect gene activation. Here, we report that diverse ADs form phase-separated condensates with the Mediator coactivator. For the OCT4 and GCN4 TFs, we show that the ability to form phase-separated droplets with Mediator in vitro and the ability to activate genes in vivo are dependent on the same amino acid residues. For the estrogen receptor (ER), a ligand-dependent activator, we show that estrogen enhances phase separation with Mediator, again linking phase separation with gene activation. These results suggest that diverse TFs can interact with Mediator through the phase-separating capacity of their ADs and that formation of condensates with Mediator is involved in gene activation.

Systematic analysis of binding of transcription factors to noncoding variants.

Many sequence variants have been linked to complex human traits and diseases1, but deciphering their biological functions remains challenging, as most of them reside in noncoding DNA. Here we have systematically assessed the binding of 270 human transcription factors to 95,886 noncoding variants in the human genome using an ultra-high-throughput multiplex protein-DNA binding assay, termed single-nucleotide polymorphism evaluation by systematic evolution of ligands by exponential enrichment (SNP-SELEX). The resulting 828 million measurements of transcription factor-DNA interactions enable estimation of the relative affinity of these transcription factors to each variant in vitro and evaluation of the current methods to predict the effects of noncoding variants on transcription factor binding. We show that the position weight matrices of most transcription factors lack sufficient predictive power, whereas the support vector machine combined with the gapped k-mer representation show much improved performance, when assessed on results from independent SNP-SELEX experiments involving a new set of 61,020 sequence variants. We report highly predictive models for 94 human transcription factors and demonstrate their utility in genome-wide association studies and understanding of the molecular pathways involved in diverse human traits and diseases.

EPSTI1 promotes osteoclast differentiation and bone resorption by PKR/NF-κB signaling.

BACKGROUND: Epithelial stromal interaction 1 (EPSTI1) plays an important role in M1 macrophages, which induce osteoclastogenesis. One recent genome-wide association study (GWAS) involving 426,824 individuals has shown that EPSTI1 is strongly associated with osteoporosis (P < 5E-8). Therefore, we speculate that EPSTI1 participates in the modulation of osteoporosis through osteoclastogenesis. The roles of EPSTI1 in osteoclastogenesis and bone resorption remain unclear. METHODS: Femur specimens were collected from osteoporotic patients and control patients. Immunofluorescence staining was used to detect the expression of EPSTI1 and signaling pathways. The osteoclastic potential of RAW264.7 cells with Sh-EPSTI1 lentivirus infection was tested using tartrate-resistant acid phosphatase (TRAP) staining, western blotting, and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Western blotting was also used to examine signaling pathways. RESULTS: In this study, EPSTI1 was found to be significantly increased in tartrate-resistant acid phosphatase positive (ACP5+) osteoclasts of bone sections from osteoporotic patients. Next, we identified EPSTI1 as a positive regulator of osteoclastogenesis and osteoclast differentiation capability. Diminished EPSTI1 expression resulted in reduced osteoclastic resorption. Mechanistically, EPSTI1-driven osteoclastogenesis was regulated by NF-κB pathway, which was mediated by the phosphorylation of protein kinase R (p-PKR). Furthermore, EPSTI1 participating in the modulation of osteoporosis via PKR/NF-κB pathway was also verified in the bone samples of osteoporotic patients. CONCLUSIONS: Collectively, our findings suggest that EPSTI1 may regulate osteoclast differentiation and bone resorption through PKR/NF-κB pathway and in vivo experiments are needed to further verify EPSTI1 as the therapy target for osteoporosis.

Mediator complex proximal Tail subunit MED30 is critical for Mediator core stability and cardiomyocyte transcriptional network.

Dysregulation of cardiac transcription programs has been identified in patients and families with heart failure, as well as those with morphological and functional forms of congenital heart defects. Mediator is a multi-subunit complex that plays a central role in transcription initiation by integrating regulatory signals from gene-specific transcriptional activators to RNA polymerase II (Pol II). Recently, Mediator subunit 30 (MED30), a metazoan specific Mediator subunit, has been associated with Langer-Giedion syndrome (LGS) Type II and Cornelia de Lange syndrome-4 (CDLS4), characterized by several abnormalities including congenital heart defects. A point mutation in MED30 has been identified in mouse and is associated with mitochondrial cardiomyopathy. Very recent structural analyses of Mediator revealed that MED30 localizes to the proximal Tail, anchoring Head and Tail modules, thus potentially influencing stability of the Mediator core. However, in vivo cellular and physiological roles of MED30 in maintaining Mediator core integrity remain to be tested. Here, we report that deletion of MED30 in embryonic or adult cardiomyocytes caused rapid development of cardiac defects and lethality. Importantly, cardiomyocyte specific ablation of MED30 destabilized Mediator core subunits, while the kinase module was preserved, demonstrating an essential role of MED30 in stability of the overall Mediator complex. RNAseq analyses of constitutive cardiomyocyte specific Med30 knockout (cKO) embryonic hearts and inducible cardiomyocyte specific Med30 knockout (icKO) adult cardiomyocytes further revealed critical transcription networks in cardiomyocytes controlled by Mediator. Taken together, our results demonstrated that MED30 is essential for Mediator stability and transcriptional networks in both developing and adult cardiomyocytes. Our results affirm the key role of proximal Tail modular subunits in maintaining core Mediator stability in vivo.

Neratinib + fulvestrant + trastuzumab for HR-positive, HER2-negative, HER2-mutant metastatic breast cancer: outcomes and biomarker analysis from the SUMMIT trial.

BACKGROUND: HER2 mutations are targetable alterations in patients with hormone receptor-positive (HR+) metastatic breast cancer (MBC). In the SUMMIT basket study, patients with HER2-mutant MBC received neratinib monotherapy, neratinib + fulvestrant, or neratinib + fulvestrant + trastuzumab (N + F + T). We report results from 71 patients with HR+, HER2-mutant MBC, including 21 (seven in each arm) from a randomized substudy of fulvestrant versus fulvestrant + trastuzumab (F + T) versus N + F + T. PATIENTS AND METHODS: Patients with HR+ HER2-negative MBC with activating HER2 mutation(s) and prior cyclin-dependent kinase 4/6 inhibitor (CDK4/6i) therapy received N + F + T (oral neratinib 240 mg/day with loperamide prophylaxis, intramuscular fulvestrant 500 mg on days 1, 15, and 29 of cycle 1 then q4w, intravenous trastuzumab 8 mg/kg then 6 mg/kg q3w) or F + T or fulvestrant alone. Those whose disease progressed on F + T or fulvestrant could cross-over to N + F + T. Efficacy endpoints included investigator-assessed objective response rate (ORR), clinical benefit rate (RECIST v1.1), duration of response, and progression-free survival (PFS). Plasma and/or formalin-fixed paraffin-embedded tissue samples were collected at baseline; plasma was collected during and at end of treatment. Extracted DNA was analyzed by next-generation sequencing. RESULTS: ORR for 57 N + F + T-treated patients was 39% [95% confidence interval (CI) 26% to 52%); median PFS was 8.3 months (95% CI 6.0-15.1 months). No responses occurred in fulvestrant- or F + T-treated patients; responses in patients crossing over to N + F + T supported the requirement for neratinib in the triplet. Responses were observed in patients with ductal and lobular histology, 1 or ≥1 HER2 mutations, and co-occurring HER3 mutations. Longitudinal circulating tumor DNA sequencing revealed acquisition of additional HER2 alterations, and mutations in genes including PIK3CA, enabling further precision targeting and possible re-response. CONCLUSIONS: The benefit of N + F + T for HR+ HER2-mutant MBC after progression on CDK4/6is is clinically meaningful and, based on this study, N + F + T has been included in the National Comprehensive Cancer Network treatment guidelines. SUMMIT has improved our understanding of the translational implications of targeting HER2 mutations with neratinib-based therapy.

Clinical Comparison of 3 Different Double Eyelid Surgeries.

OBJECTIVE: Double blepharoplasty or double eyelid surgery is a common plastic surgery in Asians. Our study aimed to evaluate the advantages and disadvantages of the minimally invasive 3-point subcutaneous tunnel double eyelid surgical technique relative to other commonly used approaches. METHOD: The study compared 852 single eyelid patients electing 1 of 3 double eyelid surgical options; minimally invasive 3-point subcutaneous tunnel, minimally invasive 3-point embedding, or incision double eyelid surgery. Evaluation criteria included postoperative swelling, hyperemia, pain, scarring, single eyelid recurrence, and patient satisfaction after 2 years. RESULTS: All surgical methods effectively achieved double eyelids. Directly after the procedure, the minimally invasive 3-point embedding method produced the mildest degree of swelling and congestion, followed by the 3-point subcutaneous tunnel method and then the incision method (F = 523.908, P = 0.000). The incision method also resulted in more serious postoperative pain, whereas the other 2 methods were equivalent (F = 83.117, P = 0.000). Patients in the incision group had the most severe scars compared with the other 2 groups with milder scarring (F = 882.102, P = 0.000). The minimally invasive 3-point embedding method group had the highest single eyelid recurrence rate (5.88%), whereas the recurrence rates for the other 2 methods were less than 1%. Patients satisfaction after 2 years was highest in the minimally invasive 3-point subcutaneous tunneling method, followed by the 3-point embedding method and lastly the incision method (F = 25.931, P = 0.00). CONCLUSIONS: Minimally invasive 3-point subcutaneous tunnel double eyelid surgery has advantages over other techniques, and its clinical application is worth promoting for suitable patients.

Analysis of the Characteristics of Aging Changes in the Eye Contours of Chinese Han Women.

OBJECTIVE: To explore the characteristics of aging changes in the eye contour of Chinese Han women and to provide reference data to base eye rejuvenation treatments. METHOD: A total of 527 Chinese Han women without prior cosmetic or reconstructive surgery were grouped according to decades of age (123 individuals from 20 to 29 years, 104 from 30to 39 years, 100 from 40 to 49 years, 100 from 50 to 59 years, and 100 >60 years). Unobscured facial images of each individual were taken and used to measure eye contour data according to a defined matrix. The relationship between eye contour indices among the different age groups was analyzed by one-way ANOVA. RESULTS: With increasing age, the distance and the vertical distance between the medial and lateral canthal points all became progressively shorter, especially the exposed length of the eyeball. Additionally, the distance between the lateral canthus and the temporal border, the distance between the medial canthus and the lateral canthus, the distance between the lower eyelid margin and the line connecting the medial canthus and the lateral canthus, all became shorter with age. The width of the face in the lateral canthal plane became narrower, while conversely, the distance and vertical distance between the punctum and the medial canthus became longer. CONCLUSION: The eye contours of Chinese Han women show regular changes during the aging process. The main manifestations involve the cleft of the eye shrinking, the medial canthus receding, and the eye contour shifting inwards and downwards as a whole. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Efficacy of Early Application of 1540 nm Non-ablative Fractional Laser and Silicones to Improve Post-epicanthoplasty Scars.

The combination of epicanthoplasty and blepharoplasty is a popular choice in Asians to acquire a pair of charming big eyes. However, the obvious scarring in the medial canthal area may result in unsatisfactory aesthetic outcomes. Recently, various laser treatments have been used to deal with scars had achieved detectable improvement, but only a few studies investigating the efficacy of lasers used in post-epicanthoplasty scarring were developed. A total of 70 participants who underwent Z-epicanthoplasty were enrolled in this prospective clinical trial and were randomly assigned to the groups of 1540nm non-ablative fractional laser (NAFL) combined with silicones treatment and silicones alone treatment. The NAFL-exposure in the treated group was additionally applied to the medial canthal area on day 21 postoperatively, compared with the participants in the control group who had only been treated with the daily usage of silicone sheets for 12 hours and silicone gels twice for 5 months after scab had fallen from the skin. Scar evolution was performed by patient and observer scar assessment Scales (POSAS) and visual analogue scale (VAS) for 21 days, and 6 months postoperatively. 64 participants have completed the entire follow-up process. The scar recovery was statistically detected in treated group compared with the control group at 6-month postoperatively assessed by POSAS, especially in pliability of scars. Furthermore, the VAS evaluations showed superior satisfaction in treated group. The early treatment of NAFL combined with silicones has improved scar formation in medial canthal region after epicantholplasties efficiently.Level of Evidence IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Embedding Group VIII Elements into a 2D Rigid pc-C3N2 Monolayer to Achieve Single-Atom Catalysts with Excellent OER Activity: A DFT Theoretical Study.

Under DFT calculations, a systematic investigation is carried out to explore the structures and oxygen evolution reaction (OER) catalytic activities of a series of 2D single-atom catalyst (SAC) systems, which are constructed by doping the transition metal (TM) atoms in group VIII into the cavities of rigid phthalocyanine carbide (pc-C3N2). We can find that when Co, Rh, Ir and Ru atoms are doped in the small or large cavities of a pc-C3N2 monolayer, they can be used as high-activity centers of OER. All these four new TM@C3N2 nanostructures can exhibit very low overpotential values in the range of 0.33~0.48 V, even smaller than the state-of-the-art IrO2 (0.56 V), which indicates considerably high OER catalytic activity. In particular, the Rh@C3N2 system can show the best OER performance, given that doped Rh atoms can uniformly serve as high-OER-active centers, regardless of the size of cavity. In addition, a detailed mechanism analysis was carried out. It is found that in these doped pc-C3N2 systems, the number of outer electrons, the periodic number of doped TM atoms and the size of the embedded cavity can be considered the key factors affecting the OER catalytic activity, and excellent OER catalytic performance can be achieved through their effective cooperation. These fascinating findings can be advantageous for realizing low-cost and high-performance SAC catalysts for OER in the near future.

[A prospective cohort study on refractive status of schoolchildren in Huangzhong District, Xining City, Qinghai Province].

Objective: To determine the characteristics and progress of the visual acuity and refractive state of schoolchildren in Huangzhong District, Xining City, Qinghai Province in China. Methods: Cohort study. Department of Ophthalmology, Beijing Children's Hospital carried out a cohort study by collecting the visual acuity and refractive state of Grade 1-5 schoolchildren among 16 primary schools in Huangzhong District, Xining City, Qinghai Province in September 2020 and July 2021. Cycloplegic retinoscopy with eye drop which contained tropicamide (0.5%) and phenylephrine hydrochloride (0.5%) was performed in children with low vision(<1.0). Myopia was defined as the spherical equivalent (SE) ≤-0.5 D after cycloplegic retinoscopy. Measurement data was analyzed by t-test and enumeration data was analyzed by χ2 test. Multiple linear regression was used to analyze the influencing factors. Results: The 2 489 individuals with repeated tests in two years were included in the follow-up study, among whom the prevalence of myopia was 26.24%(653/2 489) in 2020, while 32.94% (820/2 489)respectively in 2021. The incidence of myopia in one school year from grades 1 to 5 was 11.19%(47/420), 5.44%(21/386), 6.39%(25/391), 11.52%(44/382) and 11.67%(30/257). The average SE of children in all grades in 2021 increased negatively from the previous year (Grade 1 to Grade 5 increased respectively: 0.40 D, 0.69 D, 0.62 D, 0.52 D and 0.37 D). Conclusions: The prevalence of myopia among schoolchildren in Huangzhong District, Xining City, Qinghai Province was relatively high. There were two peaks of myopia incidence in the first, fourth and fifth grades. Female, age, and the baseline of SE were the related influencing factors for myopia progression.

Phenotypic and genotypic characteristics of Acinetobacter baumannii enrolled in the relationship among antibiotic resistance, biofilm formation and motility.

Acinetobacter baumannii is an important pathogen in clinical. The factors of biofilm formation, antibiotic resistance and motility contribute great to A. baumannii in persisting in stressed environment, and further leads to nosocomial infections. 70 A. baumannii clinical isolates were investigated for their clinical characteristics of infection. Among the tested strains, 54 (77.1%) isolates were obtained from ICUs, with the frequency of multidrug-resistance (MDR) at 55.7%, and that of extensively drug-resistance (XDR) at 31.4%. 97.1% of the clinical isolates could form biofilms, in which 4.3% possessed weak biofilm formation ability, while 41.4% and 51.4% were moderate and strong biofilm producers, respectively. A strong correlation between antibiotic resistance and biofilm formation ability was found that all the resistant strains could form biofilms, with the majority in moderate and strong levels, but 2.9% sensitive isolates had no such ability. However, the sensitive strains that could produce biofilms showed stronger biofilm formation capacity in the early stage before 24 h compared to the resistant isolates, though they became weaker afterwards. 24 biofilm-related genes and two blaOXA genes were found in both biofilm-forming and non-biofilm-forming strains, but with higher prevalence in the strains that could produce biofilms. No correlation was detected between twitching motility with antibiotic susceptibility or biofilm formation. These results raised a viewpoint that examining timepoint is a key factor for determining the biofilm formation ability, and further highlighted the importance of the appropriate surveillance and control measures in preventing the emergence and transmission of MDR and XDR A. baumannii.

The 5-year overall survival of cervical cancer in stage IIIC-r was little different to stage I and II: a retrospective analysis from a single center.

BACKGROUND: The 2018 International Federation of Gynecology and Obstetrics (FIGO) staging guideline for cervical cancer includes stage IIIC recognized by preoperative radiology (IIIC-r) to state there are lymph nodes metastases (LNM) identified by imaging tools. We aim to explore the reasonability and limitations of stage IIIC-r and try to explore the potential reasons. METHODS: Electronic medical records were used to identify patients with cervical cancer. According to the new staging guidelines, patients were reclassified and assigned into five cohorts: stage I, stage II, stage IIIC-r, LNM confirmed by pathology (IIIC-p) and LNM detected by radiology and confirmed by pathology (IIIC r + p). Five-year overall survivals were estimated for each cohort. The diagnosis accuracy of computed tomography (CT), magnetic resonance imaging (MRI) and diameter of detected lymph nodes were also evaluated. RESULTS: A total of 619 patients were identified. The mean follow-up months were 65 months (95% CI 64.43-65.77) for all patients. By comparison, the 5-year overall survival rates were not statistically different (p = 0.21) among stage IIIC-r, stage I and stage II. While, the rates were both statistical different (p<0.001) among stage IIIC-p, IIIC r + p and stage I and stage II. The sensitivities of CT and MRI in detecting LNM preoperatively were 51.2 and 48.8%. The mean maximum diameter of pelvic lymph nodes detected by CT cohort was 1.2 cm in IIIC-r cohort, and was 1.3 cm in IIIC r + p cohort. While, the mean maximum diameter of pelvic lymph nodes detected by MRI was 1.2 cm in IIIC-r cohort, and was 1.48 cm in IIIC r + p cohort. When the diagnosis efficacy of the diameter of pelvic lymph nodes in detecting LNM were evaluated, the area under the receiver operating characteristic curve (ROC curve) was 0.58 (p = 0.05). CONCLUSIONS: It seems that the FIGO 2018 staging guideline for cervical cancer is likely to has certain limitations for the classification of those with LNM. CT or MRI, however, has limitations on detecting LNM. It would be better to use more accurate imaging tools to identify LNM in the clinical practices.

Long non-coding RNA OIP5-AS1 serves as a competing endogenous RNA to modulate X-linked inhibitor of apoptosis protein expression via adsorbing miR-429 in papillary thyroid cancer.

Papillary thyroid cancer (PTC) is currently one of the most common endocrine tumors worldwide. Long non-coding RNA (LncRNA) is a vital regulator in the biological processes of diverse tumors. Hence, this work aimed to clarify the role and mechanism of lncRNA OIP5-AS1 in PTC progression. OIP5-AS1 and miR-429 expression levels in PTC tissues and cells were examined using qRT-PCR. Immunohistochemical staining (IHC) was applied to detect X-linked inhibitors of apoptosis protein (XIAP) expression in PTC tissues. A dual-luciferase reporter gene experiment was employed to validate the relationship for miR-429 and XIAP, miR-429 and OIP5-AS1. The regulatory effects of OIP5-AS1 on PTC cell proliferation, migration, and invasion was detected using the MTT, BrdU, Transwell and Western blot assays. In this work we reported that OIP5-AS1 expression was up-modulated in PTC tissues and cell lines. OIP5-AS1 overexpression enhanced the proliferation and metastasis of PTC cells, but the transfection of miR-429 mimics reversed the functions of OIP5-AS1 on the proliferation, migration, and invasion of PTC cells. Additionally, OIP5-AS1 was identified as a competitive endogenous RNA (ceRNA) that repressed miR-429, thereby increasing the expression level of XIAP. Taken together, the findings confirm that OIP5-AS1 accelerates PTC progression via modulating the miR-429/XIAP axis and imply that OIP5-AS1 is likely to be a therapeutic target for PTC.

Reliability and Validity of the Chinese Version of the Lymphedema Quality of Life Questionnaire.

Objective To study the reliability and validity of the Chinese version of the Lymphedema Quality of Life Questionnaire (LYMQOL) in lymphedema patients. Methods LYMQOL was translated into Chinese. The Chinese version of the LYMQOL was distributed with the official Wechat account "Lymphedema Channel" to lymphedema patients who were recruited from October 28 th, 2020 to February 23rd, 2021. Patients with upper limb lymphedema and lower limb lymphedema completed the LYMQOL-ARM subscale and the LYMQOL-LEG subscale separately, at enrollment, 1 week later, and 1 month later. Reliability, validity, feasibility, responsiveness and average time required for completing the questionnaire were assessed. Results A total of 195 patients participated in the study. The Chinese questionnaire showed high reliability with Cronbach's α coefficients of 0.849-0.902 for the LYMQOL-ARM and intraclass correlation coefficient (ICC) of 0.848-0.884 and Cronbach's α coefficients of 0.726-0.902 for the LYMQOL-LEG and ICC of 0.863-0.900. The LYMQOL showed moderate to good correlations with the EQ-5D (0.4

Factors Influencing Hypertrophic Scarring after Thyroidectomy.

OBJECTIVE: To identify the risk factors of hypertrophic scarring (HS) after thyroidectomy and construct a risk prediction model. METHODS: From November 2018 to March 2019, the clinical data of patients undergoing thyroidectomy were collected for retrospective analysis. According to the occurrence of HS, the patients were divided into an HS group and a non-HS group. Univariate analysis and binary logistic regression analysis were conducted to explore the independent risk factors for HS. Receiver operating characteristic analysis was also carried out. RESULTS: In this sample, 121 of 385 patients developed HS, an incidence of 31.4%. Univariate analysis showed significant differences in sex, age, postoperative infection, history of abnormal wound healing, history of pathologic scar, family history of pathologic scar, and scar prevention measures between the two groups (P < .05). Binary logistic regression analysis indicated that age 45 years or younger (odds ratio [OR], 1.815), history of abnormal wound healing (OR, 4.247), history of pathologic scarring (OR, 9.840), family history of pathologic scarring (OR, 5.708), and absence of preventive scar measures (OR, 5.566) were independent factors for HS after thyroidectomy. The area under the receiver operating characteristic curve was 0.837. When the optimal diagnostic cutoff value was 0.206, the sensitivity was 0.661, and the specificity was 0.932. CONCLUSIONS: The development of HS after thyroidectomy is related to many factors, and the proposed risk prediction model based on the combined risk factors shows a good predictive value for postoperative HS. When researchers consider the prevention and treatment of scarring in patients at risk, the incidence of HS in different populations can provide theoretical support for clinical decision-making.

CDP-DAG synthase 1 and 2 regulate lipid droplet growth through distinct mechanisms.

Lipid droplets (LDs) are evolutionarily conserved organelles that play critical roles in mammalian lipid storage and metabolism. However, the molecular mechanisms governing the biogenesis and growth of LDs remain poorly understood. Phosphatidic acid (PA) is a precursor of phospholipids and triacylglycerols and substrate of CDP-diacylglycerol (CDP-DAG) synthase 1 (CDS1) and CDS2, which catalyze the formation of CDP-DAG. Here, using siRNA-based gene knockdowns and CRISPR/Cas9-mediated gene knockouts, along with immunological, molecular, and fluorescence microscopy approaches, we examined the role of CDS1 and CDS2 in LD biogenesis and growth. Knockdown of either CDS1 or CDS2 expression resulted in the formation of giant or supersized LDs in cultured mammalian cells. Interestingly, down-regulation of cell death-inducing DFF45-like effector C (CIDEC), encoding a prominent regulator of LD growth in adipocytes, restored LD size in CDS1- but not in CDS2-deficient cells. On the other hand, reducing expression of two enzymes responsible for triacylglycerol synthesis, diacylglycerol O-acyltransferase 2 (DGAT2) and glycerol-3-phosphate acyltransferase 4 (GPAT4), rescued the LD phenotype in CDS2-deficient, but not CDS1-deficient, cells. Moreover, CDS2 deficiency, but not CDS1 deficiency, promoted the LD association of DGAT2 and GPAT4 and impaired initial LD maturation. Finally, although both CDS1 and CDS2 appeared to regulate PA levels on the LD surface, CDS2 had a stronger effect. We conclude that CDS1 and CDS2 regulate LD dynamics through distinct mechanisms.

Molecular characterization of gut microbial shift in SD rats after death for 30 days.

To observe the temporal shifts of the intestinal microbial community structure and diversity in rats for 30 days after death. Rectal swabs were collected from rats before death (BD) and on day 1, 5, 10, 15, 20, 25, and 30 after death (AD). Bacteria genomic DNA was extracted and V3 + V4 regions of 16S rRNA gene were amplified by PCR. The amplicons were sequenced at Illumina MiSeq sequencing platform. The bacterial diversity and richness showed similar results from day 1 to 5 and day 10 to 25 all presenting downtrend, while from day 5 to 10 showed slightly increased. The relative abundance of Firmicutes and Proteobacteria displayed inverse variation in day 1, 5, 10 and that was the former decreased, the latter increased. Bacteroidetes, Spirochaete and TM7 in day 15, 20, 25, 30 was significantly decline comparing with BD. Enterococcus and Proteus displayed reduced trend over day 1, 5, 10 and day 10, 15, 20, 25, respectively, while Sporosarcina showed obvious elevation during day 15, 20, 25. Accordingly, there was a certain correlation between intestinal flora succession and the time of death. The results suggested that intestinal flora may be potential indicator to aid estimation of post-mortem interval (PMI).

Individualized Surgical Treatment of Different Types of Labia Minora Hypertrophy.

Objective To explore an optimal surgical approach for different types of labia minora hypertrophy and to design a better personalized surgical treatment plan for patients. Methods Forty-five patients with labia minora treated in our department from January 2014 to January 2019 were the study participants. Depending upon the appearance of the labia minora, they were divided into length labia minora hypertrophy, width labia minora hypertrophy, overall labia minora hypertrophy, and labia hypertrophy combined with clitoris foreskin. By combining the characteristics of the patient's labia minora hypertrophy and the aesthetic requirements for postoperative appearance, we designed a personalized treatment plan for each group of patients. The surgical approaches include upper pedicle flap wedge resection, upper and lower pedicle flap similar to wedge resection, lower pedicle flap pedicle wedge resection, and labia minora combined with clitoris foreskin surgery. We followed up the postoperative results, recovery, and complication rates for a period of 3 months. Results All patients who underwent surgery had good wound healing. The labia minora had good bilateral symmetry, moderate size, and desired appearance. The scar of the surgical incision was also not obvious. The postoperative labia minora felt normal without any serious complications. Overall, the treatment effect was satisfactory. Conclusion There are many surgical methods for the reduction in the labia minora. Our findings suggest that in order to achieve better results, a personalized surgical plan should be designed considering the patient's unique type of hypertrophy and the individual aesthetic requirements. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Pretreatment thrombocytosis predict poor prognosis in patients with endometrial carcinoma: a systematic review and meta-analysis.

BACKGROUND: Several previous studies have confirmed that thrombocytosis was related to reduced survival in many solid tumors. However, the prognostic significance of thrombocytosis in endometrial carcinoma (EC) was still controversy. Therefore, we conducted this study to assess the prognostic value of thrombocytosis in EC. METHODS: The database including PubMed, MEDLINE, EMBASE, and Web of Science was searched to explore available literature. Above all, the hazard ratio (HR), odds ratios (OR) with 95% confidence intervals (CIs) was used to investigate the correlation between thrombocytosis and overall survival (OS) and disease-free survival (DFS). Moreover, the association between thrombocytosis and patient clinicopathological characteristics was explored. Publication bias and sensitivity analysis also were conducted in this study. RESULTS: Overall, 11 studies involving 3439 patients were contained in this study. The results revealed that pretreatment thrombocytosis was significantly related to a decreased OS (pooled HR = 2.99; 95% CI = 2.35-3.8; P < 0.001) and DFS (pooled HR = 2.86; 95% CI = 2.27-3.6; P <  0.001) in patients with EC. Moreover, thrombocytosis was correlated with adverse clinicopathological parameters. CONCLUSIONS: Pretreatment thrombocytosis is an adverse prognostic marker in patients with EC.